BD FACSCanto II, Reference Manual, Page: 118 / 158

118 BD FACSCanto II Flow Cytometer Reference Manual
The green circle represents a population stained with FITC. The gray circle
represents a population negative for both FITC and PE.
Figure A-12 Theoretical display of FITC vs PE before compensation
If the fluorescence is to be assigned to PE, the FITC signal must be removed from
the PE channel, as indicated by the arrow. Both PE and FITC fluoresce in the
yellow (575 nm) range, so there is no way to isolate the emission from each
fluorochrome optically. Instead, fluorescence compensation moves the FITC
population out of the PE positive area (Figure A-13).
Figure A-13 FITC signal properly compensated out of the PE channel
The software automatically determines these compensations during setup, which
you can also manually adjust:
• In BD FACSCanto clinical software, adjust spectral overlap on the Spectral
Overlap tab.
• In BD FACSDiva software, adjust spectral overlap in the Compensation tab
in the Instrument frame.
PE
FITC
PE
FITC